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. 1991 Dec;173(24):7867–7874. doi: 10.1128/jb.173.24.7867-7874.1991

Effect of chromosome location of Bacillus subtilis forespore genes on their spo gene dependence and transcription by E sigma F: identification of features of good E sigma F-dependent promoters.

D Sun 1, P Fajardo-Cavazos 1, M D Sussman 1, F Tovar-Rojo 1, R M Cabrera-Martinez 1, P Setlow 1
PMCID: PMC212578  PMID: 1744043

Abstract

Translational lacZ fusions to forespore genes of Bacillus subtilis were not expressed in spoIIAC (sigma F) or spoIIIE mutants when the lacZ fusions were integrated at the loci of the same genes or at the SP beta locus. However, some of these genes, including gerA, gpr, spoIIIG (sigma G), and sspE, were expressed in spoIIIE mutants and spoIIIE spoIIIG double mutants (but not in spoIIAC mutants) when the lacZ fusions were integrated at the amyE locus. When tested, the beta-galactosidase made in these mutants was found only in the forespore, and the 5' ends of the mRNAs produced in these mutants were identical to those in a Spo+ background. Analysis of the in vitro transcription of forespore genes by RNA polymerase containing sigma F (E sigma F) revealed a direct correlation between good in vitro transcription by E sigma F and expression at the amyE locus in spoIIIE mutants. This result suggests that forespore genes are transcribed by E sigma F in spoIIIE and spoIIIE spoIIIG mutants. Comparison of the promoter regions of genes transcribed well and poorly by E sigma F in vivo and in vitro showed that good transcription by E sigma F was correlated with G residues at positions -15 and -16, a purine residue at position -13, and a T residue at position -7 relative to the start site of transcription. The importance of these residues in sigma F recognition was confirmed by analysis of the E sigma F-dependent transcription in vivo and in vitro of mutant ssp genes.

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Selected References

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  1. Amjad M., Castro J. M., Sandoval H., Wu J. J., Yang M., Henner D. J., Piggot P. J. An SfiI restriction map of the Bacillus subtilis 168 genome. Gene. 1991 May 15;101(1):15–21. doi: 10.1016/0378-1119(91)90219-2. [DOI] [PubMed] [Google Scholar]
  2. Connors M. J., Mason J. M., Setlow P. Cloning and nucleotide sequencing of genes for three small, acid-soluble proteins from Bacillus subtilis spores. J Bacteriol. 1986 May;166(2):417–425. doi: 10.1128/jb.166.2.417-425.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fajardo-Cavazos P., Tovar-Rojo F., Setlow P. Effect of promoter mutations and upstream deletions on the expression of genes coding for small, acid-soluble spore proteins of Bacillus subtilis. J Bacteriol. 1991 Mar;173(6):2011–2016. doi: 10.1128/jb.173.6.2011-2016.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Feavers I. M., Foulkes J., Setlow B., Sun D., Nicholson W., Setlow P., Moir A. The regulation of transcription of the gerA spore germination operon of Bacillus subtilis. Mol Microbiol. 1990 Feb;4(2):275–282. doi: 10.1111/j.1365-2958.1990.tb00594.x. [DOI] [PubMed] [Google Scholar]
  5. Foulger D., Errington J. The role of the sporulation gene spoIIIE in the regulation of prespore-specific gene expression in Bacillus subtilis. Mol Microbiol. 1989 Sep;3(9):1247–1255. doi: 10.1111/j.1365-2958.1989.tb00275.x. [DOI] [PubMed] [Google Scholar]
  6. Francesconi S. C., MacAlister T. J., Setlow B., Setlow P. Immunoelectron microscopic localization of small, acid-soluble spore proteins in sporulating cells of Bacillus subtilis. J Bacteriol. 1988 Dec;170(12):5963–5967. doi: 10.1128/jb.170.12.5963-5967.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goldrick S., Setlow P. Expression of a Bacillus megaterium sporulation-specific gene during sporulation of Bacillus subtilis. J Bacteriol. 1983 Sep;155(3):1459–1462. doi: 10.1128/jb.155.3.1459-1462.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Graves M. C., Rabinowitz J. C. In vivo and in vitro transcription of the Clostridium pasteurianum ferredoxin gene. Evidence for "extended" promoter elements in gram-positive organisms. J Biol Chem. 1986 Aug 25;261(24):11409–11415. [PubMed] [Google Scholar]
  9. Hackett R. H., Setlow P. Cloning, nucleotide sequencing, and genetic mapping of the gene for small, acid-soluble spore protein gamma of Bacillus subtilis. J Bacteriol. 1987 May;169(5):1985–1992. doi: 10.1128/jb.169.5.1985-1992.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Henkin T. M., Sonenshein A. L. Mutations of the Escherichia coli lacUV5 promoter resulting in increased expression in Bacillus subtilis. Mol Gen Genet. 1987 Oct;209(3):467–474. doi: 10.1007/BF00331151. [DOI] [PubMed] [Google Scholar]
  11. Karmazyn-Campelli C., Bonamy C., Savelli B., Stragier P. Tandem genes encoding sigma-factors for consecutive steps of development in Bacillus subtilis. Genes Dev. 1989 Feb;3(2):150–157. doi: 10.1101/gad.3.2.150. [DOI] [PubMed] [Google Scholar]
  12. Kunkel B., Sandman K., Panzer S., Youngman P., Losick R. The promoter for a sporulation gene in the spoIVC locus of Bacillus subtilis and its use in studies of temporal and spatial control of gene expression. J Bacteriol. 1988 Aug;170(8):3513–3522. doi: 10.1128/jb.170.8.3513-3522.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mandelstam J., Errington J. Dependent sequences of gene expression controlling spore formation in Bacillus subtilis. Microbiol Sci. 1987 Aug;4(8):238–244. [PubMed] [Google Scholar]
  14. Mason J. M., Fajardo-Cavazos P., Setlow P. Levels of mRNAs which code for small, acid-soluble spore proteins and their LacZ gene fusions in sporulating cells of Bacillus subtilis. Nucleic Acids Res. 1988 Jul 25;16(14A):6567–6583. doi: 10.1093/nar/16.14.6567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mason J. M., Hackett R. H., Setlow P. Regulation of expression of genes coding for small, acid-soluble proteins of Bacillus subtilis spores: studies using lacZ gene fusions. J Bacteriol. 1988 Jan;170(1):239–244. doi: 10.1128/jb.170.1.239-244.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nakatani Y., Nicholson W. L., Neitzke K. D., Setlow P., Freese E. Sigma-G RNA polymerase controls forespore-specific expression of the glucose dehydrogenase operon in Bacillus subtilis. Nucleic Acids Res. 1989 Feb 11;17(3):999–1017. doi: 10.1093/nar/17.3.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nicholson W. L., Sun D. X., Setlow B., Setlow P. Promoter specificity of sigma G-containing RNA polymerase from sporulating cells of Bacillus subtilis: identification of a group of forespore-specific promoters. J Bacteriol. 1989 May;171(5):2708–2718. doi: 10.1128/jb.171.5.2708-2718.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Partridge S. R., Foulger D., Errington J. The role of sigma F in prespore-specific transcription in Bacillus subtilis. Mol Microbiol. 1991 Mar;5(3):757–767. doi: 10.1111/j.1365-2958.1991.tb00746.x. [DOI] [PubMed] [Google Scholar]
  19. Piggot P. J. Mapping of asporogenous mutations of Bacillus subtilis: a minimum estimate of the number of sporeulation operons. J Bacteriol. 1973 Jun;114(3):1241–1253. doi: 10.1128/jb.114.3.1241-1253.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schmidt R., Margolis P., Duncan L., Coppolecchia R., Moran C. P., Jr, Losick R. Control of developmental transcription factor sigma F by sporulation regulatory proteins SpoIIAA and SpoIIAB in Bacillus subtilis. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9221–9225. doi: 10.1073/pnas.87.23.9221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shimotsu H., Henner D. J. Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene. 1986;43(1-2):85–94. doi: 10.1016/0378-1119(86)90011-9. [DOI] [PubMed] [Google Scholar]
  22. Sinden R. R., Pettijohn D. E. Chromosomes in living Escherichia coli cells are segregated into domains of supercoiling. Proc Natl Acad Sci U S A. 1981 Jan;78(1):224–228. doi: 10.1073/pnas.78.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sun D. X., Cabrera-Martinez R. M., Setlow P. Control of transcription of the Bacillus subtilis spoIIIG gene, which codes for the forespore-specific transcription factor sigma G. J Bacteriol. 1991 May;173(9):2977–2984. doi: 10.1128/jb.173.9.2977-2984.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sun D. X., Stragier P., Setlow P. Identification of a new sigma-factor involved in compartmentalized gene expression during sporulation of Bacillus subtilis. Genes Dev. 1989 Feb;3(2):141–149. doi: 10.1101/gad.3.2.141. [DOI] [PubMed] [Google Scholar]
  25. Sussman M. D., Setlow P. Cloning, nucleotide sequence, and regulation of the Bacillus subtilis gpr gene, which codes for the protease that initiates degradation of small, acid-soluble proteins during spore germination. J Bacteriol. 1991 Jan;173(1):291–300. doi: 10.1128/jb.173.1.291-300.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wu J. J., Howard M. G., Piggot P. J. Regulation of transcription of the Bacillus subtilis spoIIA locus. J Bacteriol. 1989 Feb;171(2):692–698. doi: 10.1128/jb.171.2.692-698.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zuber P., Losick R. Role of AbrB in Spo0A- and Spo0B-dependent utilization of a sporulation promoter in Bacillus subtilis. J Bacteriol. 1987 May;169(5):2223–2230. doi: 10.1128/jb.169.5.2223-2230.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zuberi A. R., Moir A., Feavers I. M. The nucleotide sequence and gene organization of the gerA spore germination operon of Bacillus subtilis 168. Gene. 1987;51(1):1–11. doi: 10.1016/0378-1119(87)90468-9. [DOI] [PubMed] [Google Scholar]

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